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Roles of cell adhosive peptides from laminins in organogenesis

层粘连蛋白细胞粘附肽在器官发生中的作用

基本信息

批准号：
10670028
负责人：
KADOYA Yuichi
金额：
$ 1.73万
依托单位：
KITASATO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

1. The distributions of laminin-5 and laminin-α5 chain, were studied immunohistochemically in the developing submandibular glands of mice. The expression of laminin-5 was found in the basement membrane of developing duct but not epithelial end bud. In contrast, laminin-α5 chain was expressed in the entire epithelial basement membrane at the early embryonic stage, suggesting the role of laminin-α5 chain on the epithelial morphogenesis.2. To elucidate roles of the laminin α5 chain on organogenesis, we tested 15 cell-adhesive synthetic peptides from a carboxy-terminal globular domain of laminin-α5-chain sequence in submandibular gland organ culture. We found a peptide sequence LVLFLNHGHFVA (residues 3307-3318 and designated A5G-77) strongly inhibits the organ development at 250μg/ml. Various amino- and carboxy-terminal truncated peptides of A5G-77 were synthesized and their activities were also tested in the same system. A carboxy-treminal deletion peptide A5G-77f (LVLFLNHGH) worked inhib … More itory, whereas an amino-treminal deletion peptide A5G-77c (FLNHGHFVA) failed to inhibit the organ development. The morphology of A5G-77-treated explants differed from those of AG-73-treated, indicating distinct role of individual laminin α chains for organ development.3. We established a serum-free organ culture system of isolated vibrissal rudiments of embryonic day-13 mice. This system allowed us to test 22 laminin-1 peptides for their roles on the vibrissal growth and differentiation. We found that RKRLQVQLSIRT sequence, which is mapped at LG-4 subdomain of laminin-α1-chain carboxyl-terminal G-domain and designated AG-73, perturbed vibrissal growth in vitro. Other cell-adhesive laminin-1 peptides and a control scrambled-peptide, LQQRRSVLRTKI, failed to show any significant effects on vibrissal growth. An impaired basement membrane formation, an abnormal cytoplasmic bleb formation, and an unusual basal formation of actin bundles were noted in AG-73-treated vibrissal epidermis, indicating that AG-73 signaling is involved in cytoskeletal organization. Less

1.用免疫组织化学方法研究了层粘连蛋白5（laminin-5）和层粘连蛋白α5链（laminin-α5 chain）在小鼠颌下腺发育过程中的分布。层粘连蛋白-5表达于发育中的导管基底膜，而不表达于上皮端芽。而层粘连蛋白-α5链在胚胎早期的整个上皮基底膜上均有表达，提示层粘连蛋白-α5链在上皮形态发生中的作用.为了阐明层粘连蛋白α5链在器官发生中的作用，我们在下颌下腺器官培养中检测了15种来自层粘连蛋白α5链羧基端球状结构域序列的细胞粘附性合成肽。我们发现肽序列LVLFLNHGHFVA（残基3307-3318，命名为A5 G-77）在250μg/ml时强烈抑制器官发育。合成了A5 G-77的各种氨基和羧基端截短肽，并在同一系统中测试了它们的活性。一个羧基-苏氨酸缺失肽A5 G-77 f（LVLFLNHGH）的合成与应用 ...更多信息暂时性的，而氨基-苏氨酸缺失肽A5 G-77 c（FLNHGHFVA）不能抑制器官发育。A5 G-77处理的外植体的形态与AG-73处理的外植体不同，表明单个层粘连蛋白α链对器官发育的不同作用.我们建立了一个胚胎13天小鼠触须原基的无血清器官培养体系。该系统使我们能够测试22层粘连蛋白-1肽对触须生长和分化的作用。我们发现RKRLQVQLSIRT序列定位于层粘连蛋白α1链羧基端G结构域的LG-4亚结构域，命名为AG-73，在体外干扰触须生长。其他细胞粘附层粘连蛋白-1肽和对照乱序肽LQQRRSVLRTKI，未能显示对触须生长的任何显著影响。受损的基底膜形成，异常的细胞质水泡形成，和一个不寻常的肌动蛋白束的基础形成中指出AG-73处理的触须表皮，表明AG-73信号参与细胞骨架组织。少